US6509031B1
Expired / Lapsed
US
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System for polymerizing collagen and collagen composites in situ for a tissue compatible wound sealant, delivery vehicle, binding agent and/or chemically modifiable matrix
Assignee: Texas A&M University System
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Filing: 2000-11-15
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Granted: 2003-01-21
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Expires: 2020-11-15
Abstract
Abstract This invention relates to materials and methods for crosslinking, in situ, proteins, including collagen, with peroxidase, including horseradish peroxidase, to from biocompatible semi-solid gels useful in a number of biological and food product applications.
Inventors
- 👤 Douglas R. Miller
- 👤 Ian R. Tizard
- 👤 Jimmy T. Keeton
- 👤 Jerry F. Prochaska
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:05
CPC Classifications
A; A23; A23J; A23J3/00; A23J3/04; A23J3/30; A23J3/32; A23J3/34; A23J3/341; A23J3/342; A23L; A23L13/00; A23L13/40; A23L13/42; A23L13/424; A23L13/60; A61; A61L; A61L24/00; A61L24/04; A61L24/043; A61L24/10; A61L24/102; A61L26/00; A61L26/0009; A61L26/0028; A61L26/0033; A61L26/0042; A61L26/0052; C; C12; C12Y; C12Y111/00; C12Y111/01; C12Y111/01007
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
76 words
A method of sealing a wound comprising the steps of: applying to a wound a mixture of collagen and a crosslinking agent, wherein the collagen is selected from the group consisting of Type I collagen, Type II collagen, Type III collagen and Type IV collagen and the crosslinking agent is peroxidase and hydrogen peroxide (H 2 O 2 ); crosslinking said collagen in said wound, whereby said crosslinking forms a semi-solid gel that seals said wound.
Claim 2
Independent Claim
76 words
A method of sealing a wound comprising the steps of: applying to a wound a mixture of collagen and a crosslinking agent, wherein the collagen is selected from the group consisting of Type I collagen, Type II collagen, Type III collagen and Type IV collagen and the crosslinking agent is peroxidase and hydrogen peroxide (H 2 O 2 ); crosslinking said collagen in said wound, whereby said crosslinking forms a semi-solid gel that seals said wound.
Claim 3
Dependent Claim
53 words
The method of claim 1 , wherein the crosslinking agent is selected from the group consisting of horseradish peroxidase (HRP) and hydrogen peroxide (H 2 O 2 ); or soybean peroxidase and hydrogen peroxide (H 2 O 2 ), or microbial peroxidase from Arthromyces ramosus and hydrogen peroxide (H 2 O 2 ).
Claim 4
Dependent Claim
22 words
The method of claim 2 , wherein the crosslinking agent is horseradish peroxidase (HRP) and hydrogen peroxide (H 2 O 2 ).
Claim 5
Dependent Claim
47 words
The method of claim 3 , wherein the molar ratios of collagen to horseradish peroxidase to hydrogen peroxide range from 4 to 5 collagen to 1 horseradish peroxidase to 50 hydrogen peroxide (4-5:1:50), to 4 to 5 collagen to 1 horseradish peroxidase to 200 hydrogen peroxide (4-5:1:200).
Claim 6
Dependent Claim
17 words
The method of claim 3 , wherein the collagen concentration ranges from 8 mg/ml to 12 mg/ml.
Claim 7
Dependent Claim
17 words
The method of claim 4 , wherein the collagen concentration ranges from 8 mg/ml to 12 mg/ml.
Claim 8
Dependent Claim
13 words
The method of claim 1 , wherein the collagen is Type I collagen.
Claim 9
Dependent Claim
16 words
The method of claim 7 , wherein the collagen is Type I collagen from calf skin.
Claim 10
Dependent Claim
54 words
The method of claim 1 , wherein the semi-solid gel has properties consisting of decreased solubility of matrices in hot (75° C.) sodium dodecyl sulfate (SDS) solution, and an increase in high molecular weight components observed by SDS agarose gel filtration and SDS-polyacrylamide gel electrophoresis with a concomitant decrease in lower molecular weight components.
Claim 11
Dependent Claim
20 words
The method of claim 1 , wherein the mixture applied to the wound further comprises at least one additional agent.
Claim 12
Dependent Claim
14 words
The method of claim 10 , wherein the additional agent is a delivery agent.
Claim 13
Dependent Claim
26 words
The method of claim 10 , wherein the additional agent is selected from the group consisting of proteins, vaccine antigens, adjuvants, growth factors, microbeads and drugs.
Claim 14
Dependent Claim
30 words
The method of claim 10 , wherein the additional agent is selected from the group consisting of bovine serum albumin, fibrinogen, fibronectin, fibroblast growth factor, and human placental hyaluronic acid.
Claim 15
Dependent Claim
13 words
The method of claim 10 , wherein the additional agent is biologically active.
Claim 16
Dependent Claim
16 words
The method of claim 14 , wherein the additional agent stimulates cell growth and/or synthetic activities.
Claim 17
Dependent Claim
16 words
The method of claim 10 , wherein said additional agent in said gel has biological activity.
Claim 18
Dependent Claim
14 words
The method of claim 16 , wherein the additional agent is a growth factor.
Claim 19
Dependent Claim
14 words
The method of claim 17 , wherein said growth factor is fibroblast growth factor.
Claim 20
Dependent Claim
12 words
The method of claim 10 , wherein the additional agent is fibrinogen.
Claim 21
Dependent Claim
15 words
The method of claim 19 , wherein type I collagen and fibrinogen form a copolymer.
Claim 22
Dependent Claim
13 words
The method of claim 1 , wherein the gel is poorly or non-immunogenic.
Claim 23
Dependent Claim
21 words
The method of claim 10 , wherein the additional agent comprises biodegradable microbeads containing an antimicrobial agent allowing for delayed release.
Claim 24
Dependent Claim
19 words
The method of claim 10 , wherein the wound is in bone, and the additional agent comprises phospo-amino acids.
Claim 25
Dependent Claim
18 words
The method of claim 23 , wherein the gel provides a matrix material suitable for stimulation of mineralization.
Claim 26
Dependent Claim
24 words
The method of claim 23 , wherein the gel provides a matrix suitable for fixation of an orthopedic or dental implant in said bone.
Claim 27
Independent Claim
88 words
A method of forming a semi-solid crosslinked polymer on the surface of meat or poultry tissues for use as a food binding/restructuring agent comprising the steps of crosslinking a protein with a peroxidase in the presence of peroxide, wherein the protein is selected from the group consisting of Type I collagen, Type II collagen, Type III collagen, and Type IV collagen, fibrinogen, and recombinant fibronectin engineered protein polymer, and the peroxidase is selected from the group consisting of horseradish peroxidase, soybean peroxidase, and microbial peroxidase from Arthromyces ramosus.
Claim 28
Independent Claim
88 words
A method of forming a semi-solid crosslinked polymer on the surface of meat or poultry tissues for use as a food binding/restructuring agent comprising the steps of crosslinking a protein with a peroxidase in the presence of peroxide, wherein the protein is selected from the group consisting of Type I collagen, Type II collagen, Type III collagen, and Type IV collagen, fibrinogen, and recombinant fibronectin engineered protein polymer, and the peroxidase is selected from the group consisting of horseradish peroxidase, soybean peroxidase, and microbial peroxidase from Arthromyces ramosus.
Claim 29
Dependent Claim
19 words
The method of claim 26 , wherein the protein is type I collagen and the peroxidase is horseradish peroxidase.
Claim 30
Independent Claim
35 words
A method for growing dermal fibroblasts in vitro comprising the steps of growing the fibroblasts in a peroxide crosslinked collagen polymer comprising a three dimensional matrix for the growth of said dermal fibroblasts in vitro.
Claim 31
Independent Claim
35 words
A method for growing dermal fibroblasts in vitro comprising the steps of growing the fibroblasts in a peroxide crosslinked collagen polymer comprising a three dimensional matrix for the growth of said dermal fibroblasts in vitro.
Claim 32
Dependent Claim
13 words
The method of claim 28 wherein said dermal fibroblasts are mouse 3T3 cells.
Claim 33
Dependent Claim
23 words
The method of claim 28 , wherein the collagen is Type I collagen and has been crosslinked with horseradish peroxidase and hydrogen peroxide.
References Cited (Backward Citations)
US5376375A
Cited
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